1. Field of the Invention
This invention relates to methods and compositions for treating or preventing disease comprising the administration of immune modulatory sequences. The invention further relates to the means and methods for the identification of the immune modulatory sequences for preventing or treating disease, more particularly the treatment and prevention of autoimmune disease or inflammatory diseases. The invention also relates to the treatment or prevention of disease comprising the administration of the immune modulatory sequences alone. The invention also relates to the treatment or prevention of disease comprising the administration of the immune modulatory sequences in combination with a polynucleotide encoding self-protein(s), -polypeptide(s) or -peptide(s). The invention further relates to the treatment or prevention of disease comprising the administration of the immune modulatory sequences in combination with self-molecules, such as self-lipids, self-protein(s), self-peptide(s), self-polypeptide(s), self-glycolipid(s), self-carbohydrate(s), self-glycoprotein(s), and posttranslationally-modified self-protein(s), peptide(s), polypeptide(s), or glycoprotein(s). The invention also relates to the treatment or prevention of disease comprising the administration of the immune modulatory sequences in combination with one or more additional immune modulatory therapeutics.
The present invention also relates to methods and compositions for treating diseases in a subject associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) that are present in the subject and involved in a non-physiological state. The present invention also relates to methods and compositions for preventing diseases in a subject associated with one or more self-protein(s), -polypeptide(s) or -peptide(s) that are present in the subject and involved in a non-physiological state. The invention also relates to the administration of a combined therapy comprising an immune modulatory sequence and a polynucleotide encoding a self-protein(s), -polypeptide(s) or -peptide(s) present in a non-physiological state and associated with a disease. The invention also relates to modulating an immune response to self-molecule(s) present in an animal and involved in a non-physiological state and associated with a disease. The invention is more particularly related to the methods and compositions for treating or preventing autoimmune diseases associated with one or more self-molecule(s) present in the animal in a non-physiological state such as in multiple sclerosis (MS), rheumatoid arthritis (RA), insulin dependent diabetes mellitus (IDDM), autoimmune uveitis (AU), primary biliary cirrhosis (PBC), myasthenia gravis (MG), Sjogren's syndrome, pemphigus vulgaris (PV), scleroderma, pernicious anemia, systemic lupus erythematosus (SLE) and Grave's disease. The invention is further particularly related to other diseases associated with one or more self-molecule(s) present in the animal in a non-physiological state such as osteoarthritis, spinal cord injury, peptic ulcer disease, gout, migraine headaches, hyperlipidemia and coronary artery disease.
2. Background
Autoimmune Disease
Autoimmune disease is any disease caused by adaptive immunity that becomes misdirected at healthy cells and/or tissues of the body. Autoimmune disease affects 3% of the U.S. population, and likely a similar percentage of the industrialized world population (Jacobson et al., Clin Immunol Immunopathol, 84, 223-43, 1997). Autoimmune diseases are characterized by T and B lymphocytes that aberrantly target self-molecules, including but not limited to self-lipids, self-protein(s), self-peptide(s), self-polypeptide(s), self-glycolipid(s), self-carbohydrate(s), self-glycoprotein(s), and posttranslationally-modified self-protein(s), peptide(s), polypeptide(s), or glycoprotein(s), and derivatives thereof, thereby causing injury and or malfunction of an organ, tissue, or cell-type within the body (for example, pancreas, brain, thyroid or gastrointestinal tract) to cause the clinical manifestations of the disease (Marrack et al., Nat Med, 7, 899-905, 2001). Autoimmune diseases include diseases that affect specific tissues as well as diseases that can affect multiple tissues. This may, in part, for some diseases depend on whether the autoimmune responses are directed to a self molecule antigen confined to a particular tissue or to a self molecule antigen that is widely distributed in the body. The characteristic feature of tissue-specific autoimmunity is the selective targeting of a single tissue or individual cell type. Nevertheless, certain autoimmune diseases that target ubiquitous self molecules antigens can also affect specific tissues. For example, in polymyositis the autoimmune response targets the ubiquitous protein histidyl-tRNA synthetase, yet the clinical manifestations primarily involved autoimmune destruction of muscle.
The immune system employs a highly complex mechanism designed to generate responses to protect mammals against a variety of foreign pathogens while at the same time preventing responses against self-antigens. In addition to deciding whether to respond (antigen specificity), the immune system must also choose appropriate effector functions to deal with each pathogen (effector specificity). A cell critical in mediating and regulating these effector functions is the CD4+ T cell. Furthermore, it is the elaboration of specific cytokines from CD4+ T cells that appears to be one of the major mechanisms by which T cells mediate their functions. Thus, characterizing the types of cytokines made by CD4+ T cells as well as how their secretion is controlled is extremely important in understanding how the immune response is regulated.
The characterization of cytokine production from long-term mouse CD4+ T cell clones was first published more than 10 years ago (Mosmann et al., J. Immunol., 136:2348-2357, 1986). In these studies, it was shown that CD4+ T cells produced two distinct patterns of cytokine production, which were designated T helper 1 (Th1) and T helper 2 (Th2). Th1 cells were found to selectively produce interleukin-2 (IL-2), interferon-gamma (IFN-gamma) and lymphotoxin (LT), while Th2 clones selectively produced IL-4, IL-5, IL-6, and IL-13 (Cherwinsid et al., J. Exp. Med., 169:1229-1244, 1987). Somewhat later, additional cytokines, IL-9 and IL-10, were isolated from Th2 clones (Van Snick et al., J. Exp. Med., 169:363-368, 1989) (Fiorentino et al., J. Exp. Med., 170:2081-2095, 1989). Finally, additional cytokines, such as IL-3, granulocyte macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha) were found to be secreted by both Th1 and Th2 cells.
Autoimmune disease encompasses a wide spectrum of diseases that can affect many different organs and tissues within the body as outlined in the table below. (See, e.g., Paul, W. E. (1999) Fundamental Immunology, Fourth Edition, Lippincott-Raven, New York.)
Current therapies for human autoimmune disease include glucocorticoids, cytotoxic agents, and recently developed biological therapeutics. In general, the management of human systemic autoimmune disease is empirical and unsatisfactory. For the most part, broadly immunosuppressive drugs, such as corticosteroids, are used in a wide variety of severe autoimmune and inflammatory disorders. In addition to corticosteroids, other immunosuppressive agents are used in management of the systemic autoimmune diseases. Cyclophosphamide is an alkylating agent that causes profound depletion of both T- and B-lymphocytes and impairment of cell-mediated immunity. Cyclosporine, tacrolimus, and mycophenolate mofetil are natural products with specific properties of T-lymphocyte suppression, and they have been used to treat SLE, RA and, to a limited extent, in vasculitis and myositis. These drugs are associated with significant renal toxicity. Methotrexate is also used as a “second line” agent in RA, with the goal of reducing disease progression. It is also used in polymyositis and other connective-tissue diseases. Other approaches that have been tried include monoclonal antibodies intended to block the action of cytokines or to deplete lymphocytes. (Fox, D. A. Am. J Med., 99:82-88, 1995). Treatments for MS include interferon Beta and copolymer 1, which reduce relapse rate by 20-30% and only have a modest impact on disease progression. MS is also treated with immunosuppressive agents including methylprednisolone, other steroids, methotrexate, cladribine and cyclophosphamide. These immunosuppressive agents have minimal efficacy in treating MS. Current therapy for RA utilizes agents that non-specifically suppress or modulate immune function such as methotrexate, sulfasalazine, hydroxychloroquine, leflunamide, prednisone, as well as the recently developed TNF alpha antagonists etanercept and infliximab (Moreland et al., J Rheumatol, 28, 1431-52, 2001). Etanercept and infliximab globally block TNF alpha, making patients more susceptible to death from sepsis, aggravation of chronic mycobacterial infections, and development of demyelinating events.
In the case of organ-specific autoimmunity, a number of different therapeutic approaches have been tried. Soluble protein antigens have been administered systemically to inhibit the subsequent immune response to that antigen. Such therapies include delivery of myelin basic protein, its dominant peptide, or a mixture of myelin proteins to animals with experimental autoimmune encephalomyelitis (EAE) and humans with multiple sclerosis (Brocke et al., Nature, 379, 343-6, 1996); (Critchfield et al., Science, 263, 1139-43, 1994); Weiner et al., Annu Rev Immunol, 12, 809-37, (1994); administration of type II collagen or a mixture of collagen proteins to animals with collagen-induced arthritis and humans with rheumatoid arthritis (Gumanovskaya et al., Immunology, 97, 466-73, 1999); (McKown et al., Arthritis Rheum, 42, 1204-8, 1999), (Trentham et al., Science, 261, 1727-30, 1993); delivery of insulin to animals and humans with autoimmune diabetes (Pozzilli and Gisella Cavallo, Diabetes Metab Res Rev, 16, 306-7, 2000); and delivery of S-antigen to animals and humans with autoimmune uveitis (Nussenblatt et al., Am J Ophthalmol, 123, 583-92, 1997). A problem associated with this approach is T-cell unresponsiveness induced by systemic injection of antigen. Another approach is the attempt to design rational therapeutic strategies for the systemic administration of a peptide antigen based on the specific interaction between the T-cell receptors and peptides bound to major histocompatibility (MHC) molecules. One study using the peptide approach in an animal model of diabetes resulted in the development of antibody production to the peptide, (Hurtenbach U. et al, J Exp. Med, 177:1499, 1993). Another approach is the administration of TCR peptide immunization. See, for example, (Vandenbark A A et al., Nature, 341:541, 1989). Still another approach is the induction of oral tolerance by ingestion of peptide or protein antigens. See, for example, (Weiner H L, Immmunol Today, 18:335, 1997).
Immune responses to pathogens or tumors are currently altered by delivering proteins, polypeptides, or peptides, alone or in combination with adjuvants. For example, the hepatitis B virus vaccine contains recombinant hepatitis B virus surface antigen, a non-self antigen, formulated in aluminum hydroxide, which serves as an adjuvant. This vaccine induces an immune response against hepatitis B virus surface antigen to protect against infection. An alternative approach involves delivery of an attenuated, replication deficient, and/or non-pathogenic form of a virus or bacterium, each non-self antigens, to elicit a host protective immune response against the pathogen. For example, the oral polio vaccine is composed of a live attenuated virus, a non-self antigen, which infects cells and replicates in the vaccinated individual to induce effective immunity against polio virus, a foreign or non-self antigen, without causing clinical disease. Alternatively, the inactivated polio vaccine contains an inactivated or ‘killed’ virus that is incapable of infecting or replicating, and if administered subcutaneously, to induce protective immunity against polio virus.
Mechanisms of Initiation and Propagation of Immune Responses
Inflammatory Diseases Associated With “Nonself Molecules”: Infection with microorganisms including mycoplasma, viruses, bacteria, parasites and mycobacteria leads to inflammation in target organs, and in some cases systemic inflammation. Prominent examples include bacterial septic arthritis, Lyme arthritis, infectious uveitis, and septic shock. As part of the inate immune system, inflammatory mediators such as components of the clotting cascade, bradykinins, and complement are activated and contribute to inflammation and morbidity. The immune response in infectious disease is directed against non-self molecules present in the microorganisms, including proteins, lipids, carbohydrates, and nucleic acids. Bacterial DNA containing certain motifs referred to as “CpG” motifs, defined in more detail below, are capable of initiating inflammatory responses in animal models. For example, injection of bacterial DNA or CpG motifs, both of which are non-self molecules, into synovial joints mimics many of the inflammatory signs and symptoms that characterize septic arthritis.
Inflammatory Diseases Associated With “Self Molecules”: Many human diseases are associated with acute or chronic inflammation in the absence of any known infectious etiology. In these diseases, the immune system is active, causing the affected tissues to be inflamed and abnormally infiltrated by leukocytes and lymphocytes, but there appears to be no associated infection. Examples include osteoarthritis, coronary artery disease, Alzheimer's Disease, certain forms of dermatitis, gastritis, and pneumonitis. The predominant immune response is an innate immune response, in the absence of an adaptive immune response.
Autoimmune Diseases Associated With “Self Molecules”: Dozens of autoimmune diseases have been described, including rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, diabetes mellitus, psoriasis, and many others. Like the inflammatory diseases associated with self molecules above, the immune system is active, causing the affected tissues to be inflamed and abnormally infiltrated by leukocytes and lymphocytes, and there appears to be no associated infection. Unlike the inflammatory diseases associated with self molecules, a defining characteristic of autoimmune diseases is the presence of autoantibodies and/or T cells specific for self molecules expressed by the host. The mechanisms by which self molecules are selectively targeted by the host T and B lymphocytes are obscure. Some investigators have suggested that autoimmune diseases are triggered or exacerbated by infections with microbial pathogens. Stimulation with microbial CpG sequences is associated with an increased susceptibility to the development of animal models of autoimmune diseases such as EAE (Segal et al., J. Immunology, 158:5087, 1997) and SLE (Gilkeson et al., J, immunology, 142: 1482, 1989); however, there is little evidence to support the hypothesis that CpG sequences or microbial products can themselves trigger an autoimmune disease in an otherwise healthy animal, although inflammatory diseases can be induced. For example, several important experiments using gnotobiotic systems (i.e., animals raised in a germ free environment) have demonstrated that spontaneous development of autoimmune diseases occurs without exposure to naturally occurring microbes or microbial CpGs. Examples include development of autoimmune skin and genital disease in a germfree transgenic rodent model of ankylosing spondylitis (Taurog, J Exp Med, 180:2359, 1994); and development of lupus in 2 different models of SLE (Maldonadoi et al., J Immunol, 162: 6322, 1999; Unni et al., J Rheum, 2:35, 1975). An inducible model of SLE has also been described in which a single injection of any mouse strain with the hydrocarbon oil, pristane, leads to the development of SLE, characterized by the production of characteristic autoantibodies and immune complex-mediated kidney disease. Taken together, these experimental models suggest that spontaneous and inducible autoimmune diseases can develop in the absence of exposure to microbial DNA or CpGs.
Immunostimulatory sequences (ISS): The innate immune system is regarded as the first line of defense against microbes and pathogens. One of the most potent stimulants of the innate immune system is microbial DNA, which contains immunostimulatory sequences (ISS). The activation of innate immunity by specific immune stimulatory sequences in bacterial DNA requires a core unmethylated hexameric sequence motif consisting of 5′-purine-purine-cytosine-guanine-pyrimidine-pyrimidine-3′ for stimulation in mice and 5′-purine-pyrimidine-cytosine-guanine-pyrimidine-pyrimidine-3′ for stimulation in humans (Krieg et al., Annu Rev. Immunol., 20:709-760, 2002). Bacterial DNA and synthetic oligodeoxynucleotides (ODN) containing this dinucleotide motif, referred to as “CpG” sequences, within an immune stimulatory sequence motif have the ability to stimulate B cells to proliferate and secrete IL-6, IL-10, and immunoglobulin (Krieg et al., Nature, 374:546-549, 1995; Yi et al., J. Immunol, 157:5394-5402, 1996). ISS DNA also directly activates dendritic cells, macrophages and monocytes to secrete Th1-like cytokines such as TNF-α, IL6, and IL12 and up-regulates the expression of MHC and costimulatory molecules (Klinman et al., Proc. Nat. Acad. Sci. U.S.A., 93:2879-2883, 1996; Martin-Orozco et al., Int. Immunol., 11:1111-1118, 1999; Sparwasser et al., Eur. J. Immunol., 28:2045-2054, 1998). In mice, Toll-like receptor-9 (TLR-9) has been identified as the key receptor in the recognition of CpG motifs.
In vertebrate DNA, the frequency of CpG dinucleotides is suppressed to about one quarter of the predicted value, and the C in the CpG dinucleotide is methylated approximately 80% of the time. By contrast, bacterial DNA, like synthetic ODN, the C is not preferentially methylated in the CpG dinucleotide. Thus, bacterial DNA is structurally distinct from vertebrate DNA in its greater than 20-fold increased content of unmethylated CpG motifs. Numerous studies have established the unmethylated CpG motif as the molecular pattern within bacterial DNA that activates immune cells (Krieg et al., Annu. Rev. Immunol., 20:709-760, 2002).
CpG DNA is recognized as a potent adjuvant for its ability to induce a strong antibody response and Th1-like T-cell response to such nonself antigens as hen egg lysozyme and ovalbumin (Chu et al., J. Exp. Med., 186:1623-1631, 1997; Lipford et al., Eur. J. Immunol., 27:2340-2344, 1997). Currently, CpG DNA and CpG ODN are being utilized as therapeutic vaccines in various animal models of infectious diseases, tumors, allergic diseases, and autoimmune diseases (Krieg et al., Annu. Rev. Immunol., 20:709-760, 2002). The success of CpG as a vaccine relies heavily on its effectiveness of inducing a strong Th1-like response, and in some instances, redirecting a Th2 response to a Th1 response, such as in the allergic asthma model (Kline et al., J. Immunol., 160:2555-2559, 1998; Broide et al., J. Immunol., 161:7054-7062, 1998).
There has been significant attention given to the therapeutic applications of innate immune activation by CpG DNA. The potent non-antigen specific innate immune cell activation induced by CpG DNA is sufficient to protect mice against bacterial challenge, and even to treat established infections with intracellular pathogens (Agrawal et al., Trends Mol. Med., 8:114-121, 2002). CpG DNA also induces innate immune resistance to tumors and the regression of established tumors in mice (Dow et al., J. Immunol., 163:1552-1561, 1999; Carpenter et al., Cancer Res., 59:5429-5432, 1999; Smith et al., J. Natl. Cancer Inst., 90:1146-1154, 1998). The potent Th1 adjuvant effect of CpG DNA can even override preexisting Th2 immune responses; it has been used as an adjuvant for allergy vaccines, where it induces Th1 responses to antigens in the presence of a preexisting Th2 response, leading to decreased symptoms following subsequent allergen inhalation (Van Uden et al., J. Allergy Clin. Immunol., 104:902-910, 1999).
Immunoinhibitory sequences (IIS): Inhibitors of immunostimulatory sequence oligodeoxynucleotide (ISS-ODN) have been used to inhibit the immunostimulatory activity of ISS-ODN, for example, to suppress the immunostimulatory activity of any ISS-ODN present in recombinant expression vectors particularly in the context of gene therapy, as anti-inflammatory agents for reducing host immune responses to ISS-ODN in bacteria and viruses, as autoimmune modulator in combination with autoantigen or autoantibody conjugate to inhibit ISS-ODN stimulated Th1 mediated IL-12 production, for use as an adjuvant for Th2 immune responses to extracellular antigen, and generally to shift a host immune response from a Th1 to a Th2 response. See U.S. Pat. No. 6,255,292.
Yamada et. al, J. Immunol., 169; 5590-5594, 2002, using various in vitro immune activation cell systems evaluated IIS oligodeoxynucleotides in CpG induced immune stimulation. Yamada et. al. found that suppression by IIS oligodeoxynucleotides is dominant over stimulation by oligodeoxynucleotides and it is specific for CpG-induced immune responses. They found that the most suppressive oligonucleotide sequences contained polyG or G-C rich sequences, but a specific hexamer motif was not discovered. Krieg et al., PNAS, 95; 12631-12636, 1998, found that synthetic oligonucleotides containing neutralizing motifs defined by him as CpG dinucleotide in direct repeat clusters or with a C on the 5′ side or a G on the 3′ side, could block immune activation by immunostimulatory CpG motifs. Again, a hexamer immunoinhibitory squence was not discovered. In Zeuner et al., Arthritis and Rheumatism, 46: 2219-2224, 2002, the IIS described by Kreig at al. above, was demonstrated to reduce CpG induced arthritis in an animal model. In U.S. Pat. No. 6,225,292, Raz et al. describe a specific hexamer motif designated as 5′-purine-purine-[Y]-[Z]-pyrimidine-pyrimidine-3′ where Y is any nucleotide except cytosine, and Z is any nucleotide, wherein when Y is not guanosine or inosine, Z is guanosine or inosine, which blocks the stimulatory activity of CpG immunostimulatory sequences. In each of the above examples, the IIS was demonstrated to specifically inhibit immune activation caused by stimulatory CpG sequences.
Nucleic Acid Therapy
Antisense Therapy: Antisense oligonucleotides were originally designed as complementary to specific target genes to decrease their expression (Krieg, Annu. Rev. Immunol., 20:709-760, 2002). In order to prevent the degredation of these oligonucleotides the backbones were generally modified, such as to a phosphorothioate backbone. Although in many cases the antisense oligonucleotides did suppress the expression of target genes in tissues culture cells, in vivo experiments were less successful at altering expression. Instead, many investigators found unexpectedly that some of these oligonucleotides stimulated the immune response in vivo. For example, antisense oligonucleotide against the rev gene of the human immunodeficiency virus (HIV) had an immunostimulatory effect as manifested by increased B cell proliferation and splenomegaly (Branda et al., Biochem. Pharmacol., 45:2037-2043, 1993). Although no immediate immunostimulatory sequence motif was identified from these early studies, these findings led to the eventual search for specific immunostimulatory motifs.
Gene Therapy: Polynucleotide therapeutics, including naked DNA encoding peptides and/or polypeptides, DNA formulated in precipitation- and transfection-facilitating agents, and viral vectors have been used for “gene therapy.” Gene therapy is the delivery of a polynucleotide to provide expression of a protein or peptide, to replace a defective or absent protein or peptide in the host and/or to augment a desired physiologic function. Gene therapy includes methods that result in the integration of DNA into the genome of an individual for therapeutic purposes. Examples of gene therapy include the delivery of DNA encoding clotting factors for hemophilia, adenine deaminase for severe combined immunodeficiency, low-density lipoprotein receptor for familial hypercholesterolemia, glucocerebrosidase for Gaucher's disease, α1-antitrypsin for α1-antitrypsin deficiency, alpha- or Beta-globin genes for hemoglobinopathies, and chloride channels for cystic fibrosis (Verma and Somia, Nature, 389, 239-42, 1997).
DNA immunization to treat infection: In DNA immunization a non-replicating transcription unit can provide the template for the synthesis of proteins or protein segments that induce or provide specific immune responses in the host. Injection of naked DNA promotes vaccination against a variety of microbes and tumors (Robinson and Torres, Semin Immunol, 9, 271-83, 1997). DNA vaccines encoding specific proteins, present in viruses (hepatitis B-virus, human Immunodeficiency virus, rotavirus, and influenza virus), bacteria (mycobacterium tuberculosis), and parasites (Malaria), all non-self antigens, are being developed to prevent and treat these infections (Le et al., Vaccine, 18, 1893-901, 2000); (Robinson and Pertmer, Adv Virus Res, 55, 1-74, 2000).
DNA to treat neoplasia: DNA vaccines encoding major histocompatibility antigen class I, cytokines (IL-2, IL-12 and IFN-gamma), and tumor antigens are being developed to treat neoplasia (Wlazlo and Ertl, Arch Immunol Ther Exp, 49:1-11, 2001). For example, viral DNA encoding the B cell immunoglobulin idiotype (antigen binding region) has been administered to eliminate and protect against B cell-lymphomas (Timmerman et al., Blood, 97:1370-1377, 2001).
DNA immunization to treat autoimmune disease: Others have described DNA therapies encoding immune molecules to treat autoimmune diseases. Such DNA therapies include DNA encoding the antigen-binding regions of the T cell receptor to alter levels of autoreactive T cells driving the autoimmune response (Waisman et al., Nat Med, 2:899-905, 1996) (U.S. Pat. No. 5,939,400). DNA encoding autoantigens were attached to particles and delivered by gene gun to the skin to prevent multiple sclerosis and collagen induced arthritis. (Patent WO 97/46253) (Ramshaw et al., Immunol., and Cell Bio., 75:409413, 1997) DNA encoding adhesion molecules, cytokines (TNF alpha), chemokines (C-C chemokines), and other immune molecules (Fas-ligand) have been used to treat animal models of autoimmune disease (Youssef et al., J Clin Invest, 106:361-371, 2000); (Wildbaum et al., J Clin Invest, 106:671-679, 2000); (Wildbaum et al., J Immunol, 165:5860-5866, 2000); (Wildbaum et al., J Immunol, 161:6368-7634, 1998); (Youssef et al., J Autoimmun, 13:21-9, 1999).
It is an object of the present invention to provide a method and composition for treating or preventing a disease, particularly autoimmune disease or inflammatory disease, comprising the administration of immune modulatory nucleic acids. Another object of this invention is to provide the means of identification of the immune modulatory sequences for treating disease. Yet another object of this invention is to provide the method and means of treating a disease associated with self-protein(s), -polypeptide(s), or -peptide(s) that are present and involved in a non-physiological process in an animal comprising the administration of an immune modulatory sequence in combination with a polynucleotide encoding self-protein(s), -polypeptide(s) or -peptide(s). Another object of the present invention is to provide a composition for treating or preventing a disease associated with self-protein(s), -polypeptide(s), or -peptide(s) that is present non-physiologically in an animal. The invention further relates to the treatment or prevention of disease comprising the administration of the immune modulatory nucleic acids in combination with self-molecule(s). These and other objects of this invention will be apparent from the specification as a whole.